(1) Services
The University of Toledo Flow Cytometry Core Facility will provide instrumentation and expertise to investigators, students, technicians and the surrounding community. Samples are prepared by the individual investigators who then deliver samples to the Core for flow cytometric analysis or sorting. The core can provide help and/or generation of publication-quality graphics and experimental design.
(2) Location
The Core Facility is located on the Health Science Campus in the Health Education Building, Room 251.
(3) Hours of Operation, Fees, and Scheduling
This facility will be staffed from 8:30a.m. until 5:00 p.m., Monday through Friday. The facility is managed by Sean Linkes, M.S. and may be reached by phone at 419.383.3402.
The Core Facility will operate on a Fee for Service basis which will include data acquisition, analysis and consultation if needed.
Current Flow Core Rates Rate/Hour Self Operation
University of Toledo Investigators $30 $30
Users Outside the University $30 $30
Cancellations with less than a 24-hour notice are billed for the scheduled time. Investigators are not billed for cancellations with at least 24 hour notice. Exceptions will be determined by the Core Facility manager.
Scheduling can be made by calling 419.383.3402 or by visiting the facility. No scheduling will be accepted by email.
*All users must complete a Sample Submission Form that provides Core personnel with information needed to process experiment. Forms can be found here.
(4) Sample Preparation
Investigators (students, technicians, and other users) must prepare samples in their individual laboratories. Staining will not be done at the Core Facility. If needed, Core personnel can help/advise regarding sample preparation for the various flow protocols. Contact the Core Manager with respect to appropriate controls needed.
It is recommended that all investigators consult with the Core Manager with respect to experimental design, controls, and other needs that will help meet their goals. This will help to ensure that usage of the Core Facility will be extremely beneficial to all involved.
The facility cannot accept potentially pathogenic samples for cell sorting. Should you have questions relating to human/primate or lentivirus-containing/treated samples, please consult the Core Manager. Samples labeled with radioactive materials will not be accepted at any time.
(5) Instrumentation
The Core Facility contains the following machines:
5a. BD Biosciences FACSCalibur-Analysis only
Lasers
Argon- 488nm excitation wavelength
Red Diode-635nm excitation wavelength
Features
6 detectors
Aggregate discrimination via DDM (time of flight)
Parameters Commonly Used on FACSCalibur
FSC
SSC
FITC
Alexa 488
Alexa 633
DsRED
PE
CYC
APC
PI
FL-2 W
FL-2 A
FL-3 A
FL-3 W
5b. BD Biosciences FACSAria High-Speed Cell Sorter
Lasers
Argon- 488nm
Violet-407nm
Helium-Neon-633
Features
12 detectors
Multi-well plate sorter - coming soon
Aggregate discrimination - via width discrimination (time of flight)
Aerosol Management System-for use with Lenti-virus containing samples - coming soon
High speed sort rate ~25k events or cells/sec
Sample Temperature Control
Parameters Commonly Used on FACSAria
FSC
SSC
FITC
PE
PE-Texas Red
Percp-Cy5.5
PerCP
PI
PE-Cy7
APC
APC-Cy7
DAPI
Hoechst
Cascade Blue
Pacific Blue
Alexa 405
Alexa 430
(6) Facility Rules and Regulations
General
All samples will be prepared by the individual investigator. Staining will not be performed by the Core Facility. However, the core facility will provide help with experimental design-controls, seeding of samples, sorting and analysis preparation, and general questions regarding aspects of flow cytometry. Cancellations with at least a 24-hour notice will not be charged. Cancellations that are not made within 24 hours will be billed accordingly. Special circumstances will be noted.
Samples of Pathogenic Potential
It is the policy of the Core Facility that all information about the kind of samples being run at the facility be disclosed. The facility must be aware of any samples containing pathogenic potential such as lentivirius, etc. at the time of scheduling. The aerosol management system used for containment is on order and will be installed as soon as it arrives. Users who do not disclose pertinent information relating to samples will not be allowed to use the Core Facility for a period of time. Please plan accordingly.
Human Samples
Human samples must be pre-screened for the following: HIV, Hepatitis B and Hepatitis C. The Core Facility must be notified at the time of scheduling if these samples are used. Non-screened human samples will not be sorted; no exceptions will be made. Human samples that are fixed can be analyzed.
In the case of human samples, there must be a way to identify the source of the tissue. The individual investigators who use pre-screened human tissue will provide an identification number or another means of identifying the source of the tissue (actual names cannot be used due to HIPAA regulations). The ID and other pertinent information will be known by the individual investigator and Core Facility personnel as a way to identify the source of the tissue in case of an emergency, and will be kept for future records.
Users
Investigators, students and technicians will be allowed to run the FACSCalibur themselves. Self-assist users must be trained on the FACSCalibur prior to use of the instrument. Training will be provided and consists of a 2-hour session deemed a "first training" which provides basic information relating to general flow cytometry, machine operation, and maintenance. Samples will not be brought to this training. A second 1-hour session will conclude training and will include samples and the user "driving/operating" the machine with minimal supervision from Core personnel. Only training for the FACSCalibur will be provided. The FACSAria will be operated by Core Facility personnel and certified users only.
(7) Personnel and Contact Information
Akira Takashima, M.D., Ph.D.
Professor and Chair
Director, Flow Cytometry Core Facility
Department of Medical Microbiology & Immunology
Health Education Building, Room 229
3000 Arlington Avenue, Mail Stop 1021
Toledo, OH 43614-2598
Phone: 419.383.5423
Email: Akira.Takashima@utoledo.edu
Sean Linkes, M.S.
Manager, Flow Cytometry Core Facility
Department of Medical Microbiology & Immunology
Health Education Building, Room 251
3000 Arlington Avenue, Mail Stop 1021
Toledo, OH 43614-2598
Phone: 419.383.3402
Email: Sean.Linkes@utoledo.edu
(8) Helpful Links
Andy Oberyszyn, Research Reagent Sales Specialist: AOberyszyn@BD.com 614.785.0924 or 1.800.451.4557 ext. 1677
BD Spectra viewer: http://www.bdbiosciences.com/spectra/
Purdue Cytometry Board: http://www.cyto.purdue.edu/
Great Lakes International Image and Flow Cytometry Association (GLIIFCA): http://www.gliifca.org/
International Society of Analytical Cytology: http://www.isac-net.org/
BD website introduction to flow and Diva training: http://www.bdbiosciences.com/immunocytometry_systems/support/training/online/ITF/index.shtml
Page updated: April 12, 2007